The importance of iron chelation and iron availability during PpIX-induced photodynamic therapy

This is the accepted version of the article. Please cite the published version which is available via the DOI link in this record.Background: Protoporphyrin IX (PpIX)-induced photodynamic therapy (PDT) is being utilised as a topical method of localised ablation of certain non-melanoma skin cancers and precancers. Standardised protocols have been implemented to good effect when the disease remains superficial but improvement is required to treat thicker or acrally located conditions. Concurrent administration of an iron chelator during PpIX-PDT has been demonstrated to increase cellular accumulation of PpIX by reducing its bioconversion to haem (an iron dependent process) thus increasing cell kill on subsequent irradiation. Iron however, can also play a role in reactive oxygen species (ROS) generation and limiting its availability via chemical chelation could theoretically reduce the efficacy of PpIX-PDT, so that a response less than that maximally feasible is produced. Materials and methods: The effects of iron availability and chelation on PpIX-PDT have therefore been investigated via fluorescence quantification of PpIX accumulation, single-cell gel electrophoresis (comet assay) measurement of ROS-induced DNA damage and trypan blue exclusion assessment of cell viability. Cultured human cells were utilised and incubated in standardised iron conditions with the PpIX precursor's aminolaevulinic acid (ALA) or its methyl ester (MAL) in the presence or absence of either of the iron chelating agents desferrioxamine (DFO) or hydroxypyridinone (CP94), or alternatively iron sulphate as a source of iron. Results: ALA or MAL incubation was found to significantly increase cellular PpIX accumulation pre-irradiation as anticipated and this observation correlated with both significantly increased DNA damage and reduced cellular viability following irradiation. Co-incubation with either of the iron chelators investigated (DFO or CP94) significantly increased pre-irradiation PpIX accumulation as well as DNA damage and cell death on irradiation indicating the positive effect of iron chelation on the effectiveness of PpIX-induced PDT. The opposite effects were observed however, when the cells were co-incubated with iron sulphate, with significant reductions in pre-irradiation PpIX accumulation (ALA only) and DNA damage (ALA and MAL) being recorded indicating the negative effects excessive iron can have on PpIX-PDT effectiveness. Some dark toxicity produced by iron sulphate administration in non-irradiated control groups was also observed. Conclusion: Iron chelation and availability have therefore been observed to positively and adversely affect the PpIX-PDT process respectively and it is concluded that the effects of increased PpIX accumulation pre-irradiation produced via iron chelation outweigh any limitations reduced iron availability may have on the ability of iron to catalyse ROS generation/cascades following PpIX-induced PDT. Further investigation of iron chelation within dermatological applications where enhanced PpIX-PDT treatment effects would be beneficial is therefore warranted.Duchy Health Charity Limited, Cornwall, U

Enhancement of methyl-aminolevulinate photodynamic therapy by iron chelation with CP94: an in vitro investigation and clinical dose-escalating safety study for the treatment of nodular basal cell carcinoma.

Clinical TrialAuthor version of article. The final publication is available at Springer via: http://dx.doi.org/10.1007/s00432-008-0358-6PURPOSE: Methyl-aminolevulinate (MAL) photodynamic therapy (PDT) is a cancer therapy that combines the selective accumulation of a photosensitizer in tumor tissue with visible light (and tissue oxygen) to produce reactive oxygen species. This results in cellular damage and ablation of tumor tissue. Combining iron chelators with MAL has the potential to increase the accumulation of the photosensitizer protoporphyrin IX (PpIX) by reducing its bioconversion to heme. This paper investigates this method of enhancement both in vitro and for the first time clinically for the treatment of nodular basal cell carcinoma (BCC). METHODS: Enhancement of MAL-induced PpIX accumulation by the iron chelator CP94 was quantified fluorometrically in human cultured cells (including three dermatological cell types). An open, dose-escalating, pilot study was then conducted in patients with nodular BCC, to determine the safety of this pharmacological modification. RESULTS: Large enhancements in PpIX accumulation were observed in the cultured cells when co-incubated with the iron chelator CP94. Clinically the addition of CP94 was found to be feasible and safe. In addition greater reductions in tumor depth were observed in the CP94 co-incubated tumors. CONCLUSION: Iron chelation by CP94 is an effective enhancer of MAL-induced PpIX accumulation in vitro. This method of enhancement was safely applied to a clinical PDT protocol with no unexpected adverse effects reported. Although the clinical investigation was only intended to be a small pilot to assess safety, enhancements in tumor clearance were observed both clinically and histologically when CP94 was included in the photosensitizing cream.Duchy Health Charity Limite

Clinical investigation of the novel iron-chelating agent, CP94, to enhance topical photodynamic therapy of nodular basal cell carcinoma.

Clinical TrialMulticenter StudyThis is the peer reviewed version of the article which has been published in final form at DOI: 10.1111/j.1365-2133.2008.08668.x This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.© 2008 The AuthorsJournal Compilation © 2008 British Association of DermatologistsBACKGROUND: Photodynamic therapy (PDT) involves the activation of a photosensitizer by visible light to produce activated oxygen species within target cells, resulting in their destruction. Evidence-based guidelines support the efficacy of PDT using topical 5-aminolaevulinic acid (ALA-PDT) in actinic keratoses, Bowen disease and basal cell carcinoma (BCC). Efficacy for nodular BCC appears inferior to that for superficial BCC unless prior debulking or repeat treatments are performed. Objectives The aim of this study was to assess the safety and efficacy of adding a novel iron-chelating agent, CP94 (1,2-diethyl-3-hydroxypyridin-4-one hydrochloride), to topical ALA, to temporarily increase the accumulation of the photosensitizer in the tumour. METHODS: A mixed topical formulation of ALA + increasing concentrations of CP94 was used to carry out PDT on previously biopsied nodular BCC with no prior lesion preparation using standard light delivery. The area was assessed clinically and surgically excised 6 weeks later for histological examination. RESULTS: Enhanced PDT using 40% CP94 resulted in significantly greater clearance rates in nodular BCC than with ALA-PDT alone, in our protocol of single-treatment PDT with no lesion preparation. CONCLUSIONS: The results of this study demonstrate the safe and effective use of an enhanced ALA-PDT protocol for nodular BCC using CP94, with no adverse reactions to this modification. This is the first time this formulation has been used in patients. This formulation is now the focus of further study

Drivers of the Adoption and Exclusive Use of Clean Fuel for Cooking in Sub-Saharan Africa: Learnings and Policy Considerations from Cameroon

Household air pollution (HAP) caused by the combustion of solid fuels for cooking and heating is responsible for almost 5% of the global burden of disease. In response, the World Health Organisation (WHO) has recommended the urgent need to scale the adoption of clean fuels, such as liquefied petroleum gas (LPG), in low and middle-income countries (LMICs). To understand the drivers of the adoption and exclusive use of LPG for cooking, we analysed representative survey data from 3343 peri-urban and rural households in Southwest Cameroon. Surveys used standardised tools to collect information on fuel use, socio-demographic and household characteristics and use of LPG for clean cooking. Most households reported LPG to be clean (95%) and efficient (88%), but many also perceived it to be expensive (69%) and unsafe (64%). Positive perceptions about LPG's safety (OR = 2.49, 95% CI = 2.04, 3.05), cooking speed (OR = 4.31, 95% CI = 2.62, 7.10), affordability (OR = 1.7, 95% CI = 1.38, 2.09), availability (OR = 2.17, 95% CI = 1.72, 2.73), and its ability to cook most dishes (OR = 3.79, 95% CI = 2.87, 5.01), were significantly associated with exclusive LPG use. Socio-economic status (higher education) and household wealth (higher income) were also associated with a greater likelihood of LPG adoption. Effective strategies to raise awareness around safe use of LPG and interventions to address financial barriers are needed to scale wider adoption and sustained use of LPG for clean cooking, displacing reliance on polluting solid fuels

Small farmers and sustainability: Institutional barriers to investment and innovation in the Malaysian palm oil industry in Sabah

The Malaysian palm oil industry is well known for the social, environmental and sustainability challenges associated with its rapid growth over the past ten years. Technologies exist to reduce the conflict between national development aims of economic uplift for the rural poor, on the one hand, and ecological conservation, on the other hand, by raising yields and incomes from areas already under cultivation. But the uptake of these technologies has been slow, particularly in the smallholder sector. In this paper we explore the societal and institutional challenges that influence the investment and innovation decisions of micro and small enterprise (MSE) palm oil smallholders in Sabah, Malaysia. Based on interviews with 38 smallholders, we identify a number of factors that reduce the smallholders' propensity to invest in more sustainable practices. We discuss why more effective practices and innovations are not being adopted using the concepts of, firstly, institutional logics to explore the internal dynamics of smallholder production systems, including attitudes to sustainability and innovation; and, secondly, institutional context to explore the pressures the smallholders face, including problems of access to land, labour, capital, knowledge and technical resources. These factors include limited access to global market information, corruption and uncertainties of legal title, weak economic status and social exclusion. In discussing these factors we seek to contribute to wider theoretical debates about the factors that block innovation and investment in business improvements in marginal regions and in marginalised groups

An original phylogenetic approach identified mitochondrial haplogroup T1a1 as inversely associated with breast cancer risk in BRCA2 mutation carriers

Introduction: Individuals carrying pathogenic mutations in the BRCA1 and BRCA2 genes have a high lifetime risk of breast cancer. BRCA1 and BRCA2 are involved in DNA double-strand break repair, DNA alterations that can be caused by exposure to reactive oxygen species, a main source of which are mitochondria. Mitochondrial genome variations affect electron transport chain efficiency and reactive oxygen species production. Individuals with different mitochondrial haplogroups differ in their metabolism and sensitivity to oxidative stress. Variability in mitochondrial genetic background can alter reactive oxygen species production, leading to cancer risk. In the present study, we tested the hypothesis that mitochondrial haplogroups modify breast cancer risk in BRCA1/2 mutation carriers. Methods: We genotyped 22,214 (11,421 affected, 10,793 unaffected) mutation carriers belonging to the Consortium of Investigators of Modifiers of BRCA1/2 for 129 mitochondrial polymorphisms using the iCOGS array. Haplogroup inference and association detection were performed using a phylogenetic approach. ALTree was applied to explore the reference mitochondrial evolutionary tree and detect subclades enriched in affected or unaffected individuals. Results: We discovered that subclade T1a1 was depleted in affected BRCA2 mutation carriers compared with the rest of clade T (hazard ratio (HR) = 0.55; 95% confidence interval (CI), 0.34 to 0.88; P = 0.01). Compared with the most frequent haplogroup in the general population (that is, H and T clades), the T1a1 haplogroup has a HR of 0.62 (95% CI, 0.40 to 0.95; P = 0.03). We also identified three potential susceptibility loci, including G13708A/rs28359178, which has demonstrated an inverse association with familial breast cancer risk. Conclusions: This study illustrates how original approaches such as the phylogeny-based method we used can empower classical molecular epidemiological studies aimed at identifying association or risk modification effects.Peer reviewe

Drivers of the Adoption and Exclusive Use of Clean Fuel for Cooking in Sub-Saharan Africa: Learnings and Policy Considerations from Cameroon

Household air pollution (HAP) caused by the combustion of solid fuels for cooking and heating is responsible for almost 5% of the global burden of disease. In response, the World Health Organisation (WHO) has recommended the urgent need to scale the adoption of clean fuels, such as liquefied petroleum gas (LPG), in low and middle-income countries (LMICs). To understand the drivers of the adoption and exclusive use of LPG for cooking, we analysed representative survey data from 3343 peri-urban and rural households in Southwest Cameroon. Surveys used standardised tools to collect information on fuel use, socio-demographic and household characteristics and use of LPG for clean cooking. Most households reported LPG to be clean (95%) and efficient (88%), but many also perceived it to be expensive (69%) and unsafe (64%). Positive perceptions about LPG’s safety (OR = 2.49, 95% CI = 2.04, 3.05), cooking speed (OR = 4.31, 95% CI = 2.62, 7.10), affordability (OR = 1.7, 95% CI = 1.38, 2.09), availability (OR = 2.17, 95% CI = 1.72, 2.73), and its ability to cook most dishes (OR = 3.79, 95% CI = 2.87, 5.01), were significantly associated with exclusive LPG use. Socio-economic status (higher education) and household wealth (higher income) were also associated with a greater likelihood of LPG adoption. Effective strategies to raise awareness around safe use of LPG and interventions to address financial barriers are needed to scale wider adoption and sustained use of LPG for clean cooking, displacing reliance on polluting solid fuels